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- Title
Faster quantitative real-time PCR protocols may lose sensitivity and show increased variability.
- Authors
Hilscher, Chelsey; Vahrson, Wolfgang; Dittmer, Dirk P
- Abstract
Quantitative real-time PCR has become the method of choice for measuring mRNA transcription. Recently, fast PCR protocols have been developed as a means to increase assay throughput. Yet it is unclear whether more rapid cycling conditions preserve the original assay performance characteristics. We compared 16 primer sets directed against Epstein-Barr virus (EBV) mRNAs using universal and fast PCR cycling conditions. These primers are of clinical relevance, since they can be used to monitor viral oncogene and drug-resistance gene expression in transplant patients and EBV-associated cancers. While none of the primers failed under fast PCR conditions, the fast PCR protocols performed worse than universal cycling conditions. Fast PCR was associated with a loss of sensitivity as well as higher variability, but not with a loss of specificity or with a higher false positive rate.
- Publication
Nucleic acids research, 2005, Vol 33, Issue 21, pe182
- ISSN
1362-4962
- Publication type
Journal Article
- DOI
10.1093/nar/gni181